Gene-breaking transposon mutagenesis reveals an essential role for histone H2afza in zebrafish larval development.

Gene-breaking transposon mutagenesis reveals an essential role for histone H2afza in zebrafish larval development. Research Abstract Details 

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Gene-breaking transposon mutagenesis reveals an essential role for histone H2afza in zebrafish larval development. Abstract Text:

Sridhar Sivasubbu,Darius Balciunas,Ann E Davidson,Michael A Pickart,Spencer B Hermanson,Kirk J Wangensteen,Daniel C Wolbrink,Stephen C Ekker,

We report a novel gene tagging, identification and mutagenicity ('gene-breaking') method for the zebrafish, Danio rerio. This modular approach consists of two distinct and separable molecular cassettes. The first is a gene-finding cassette. In this study, we employed a 3' gene-tagging approach that selectively 'traps' transcripts regardless of expression status, and we show that this cassette identifies both known and novel endogenous transcripts in transgenic zebrafish. The second is a transcriptional termination mutagenicity cassette assembled from a combination of a splice acceptor and polyadenylation signal to disrupt tagged transcripts upon integration into intronic sequence. We identified both novel and conserved loci as linked phenotypic mutations using this gene-breaking strategy, generating molecularly null mutations in both larval lethal and adult viable loci. We show that the Histone 2a family member z (H2afza) variant is essential for larval development through the generation of a lethal locus with a truncation of conserved carboxy-terminal residues in the protein. In principle this gene-breaking strategy is scalable for functional genomics screens and can be used in Sleeping Beauty transposon and other gene delivery systems in the zebrafish.

Gene-breaking transposon mutagenesis reveals an essential role for histone H2afza in zebrafish larval development. Publishing Authors By Initials

S Sivasubbu,D Balciunas,AE Davidson,MA Pickart,SB Hermanson,KJ Wangensteen,DC Wolbrink,SC Ekker,

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Gene-breaking transposon mutagenesis reveals an essential role for histone H2afza in zebrafish larval development. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Mechanisms of development

VOLUME: 123

Page Numbers: 513-29

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ISSN: 0925-4773

DAY: 9

MONTH: 06

YEAR: 2006

Gene-breaking transposon mutagenesis reveals an essential role for histone H2afza in zebrafish larval development. Information

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LANGUAGE: eng

NlmUniqueID: 9101218

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Grant and Affiliation Information for Gene-breaking transposon mutagenesis reveals an essential role for histone H2afza in zebrafish larval development.

AFFILIATION: University of Minnesota, Department of Genetics, Cell Biology and Development, Arnold and Mabel Beckman Center for Transposon Research, 321 Church St SE, 6-160 Jackson Hall, Minneapolis, MN 55455, USA.

Country: Ireland

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MEDLINETA: Mech Dev

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